The space occupied by evolutionarily advanced ant societies can be subdivided into functional sites, such as broodchambers; peripheral nest chambers; kitchen middens; and foraging routes. Many predators and social parasites are specially adapted to make their living inside specific niches created by ants. In particular, the foraging paths of certain ant species are frequented by predatory and kleptoparasitic arthropods, including one striking example, the nitidulid beetle, Amphotis marginata. Adults of this species obtain the majority of their nutrition by acting as a kind of “highwayman” on the foraging trails of the ant Lasius fuliginosus, where they solicit regurgitation from food laden ant-workers by mimicking the ant’s food-begging signals. Employing food labeled with the radio isotope [superscript 32]P, we assessed the quantities of food the beetles siphoned-off of food-laden ants, and we investigated the site preferences, behavioral mechanisms and possible morphological adaptations underlying the food kleptoparasitism of A. marginata.

The membrane proximal region (MPR, residues 649–683) and transmembrane domain (TMD, residues 684–705) of the gp41 subunit of HIV-1’s envelope protein are highly conserved and are important in viral mucosal transmission, virus attachment and membrane fusion with target cells. Several structures of the trimeric membrane proximal external region (residues 662–683) of MPR have been reported at the atomic level; however, the atomic structure of the TMD still remains unknown. To elucidate the structure of both MPR and TMD, we expressed the region spanning both domains, MPR-TM (residues 649–705), in Escherichia coli as a fusion protein with maltose binding protein (MBP). MPR-TM was initially fused to the C-terminus of MBP via a 42 aa-long linker containing a TEV protease recognition site (MBP-linker-MPR-TM).

Biophysical characterization indicated that the purified MBP-linker-MPR-TM protein was a monodisperse and stable candidate for crystallization. However, crystals of the MBP-linker-MPR-TM protein could not be obtained in extensive crystallization screens. It is possible that the 42 residue-long linker between MBP and MPR-TM was interfering with crystal formation. To test this hypothesis, the 42 residue-long linker was replaced with three alanine residues. The fusion protein, MBP-AAA-MPR-TM, was similarly purified and characterized. Significantly, both the MBP-linker-MPR-TM and MBP-AAA-MPR-TM proteins strongly interacted with broadly neutralizing monoclonal antibodies 2F5 and 4E10. With epitopes accessible to the broadly neutralizing antibodies, these MBP/MPR-TM recombinant proteins may be in immunologically relevant conformations that mimic a pre-hairpin intermediate of gp41.